Anti-ACOX1/AOX antibody [EPR19038]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast) cell linelabeling ACOX1/AOX with ab184032 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR19038] - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ACOX1/AOX with ab184032 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [EPR19038] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab184032 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling ACOX1/AOX with ab184032 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on C6 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [EPR19038] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab184032 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

ACOX1/AOX was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab184032 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184032 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : NIH/3T3 whole cell lysate 10μg (Input).
Lane 2 : ab184032 IP in NIH/3T3 whole cell lysate.
Lane 3 : Rabbit IgG, monoclonal [EPR19038] - Isotype Control (ab172730) instead of ab184032 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-ACOX1/AOX antibody [EPR19038] (ab184032)

Predicted band size: 74 kDa

false

• WB

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Western blot - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

Blocking/dilution buffer : 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID : ) 8798738 and 17255948.

All lanes:

Western blot - Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/1000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 74 kDa

Observed band size: 50 kDa,72 kDa

false

Exposure time: 3min

• WB

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Western blot - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

Blocking/dilution buffer : 5% NFDM/TBST.

Exposure time Lane 1-5 : 1 second; Lane 6 : 30 seconds; Lane 7 : 2 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : ) 8798738 and 17255948.

Lanes 1 - 5:

Western blot - Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/1000 dilution

Lanes 6 - 7:

Western blot - Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/5000 dilution

Lane 1:

Mouse liver lysate at 10 µg

Lane 2:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 3:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 4:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 5:

NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 10 µg

Lane 6:

Mouse heart lysate at 10 µg

Lane 7:

Mouse kidney lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 74 kDa

Observed band size: 50 kDa,72 kDa

false

• WB

Supplier Data

Western blot - Anti-ACOX1/AOX antibody [EPR19038] (AB184032)

Blocking/dilution buffer : 5% NFDM/TBST.

Exposure time Lane 1/2 : 8 seconds;Lane 3 : 1 second.

The expression profile observed is consistent with what has been described in the literature (PMID : ) 8798738 and 17255948.

All lanes:

Western blot - Anti-ACOX1/AOX antibody [EPR19038] (ab184032) at 1/1000 dilution

Lane 1:

Rat brain lysate at 10 µg

Lane 2:

Rat heart lysate at 10 µg

Lane 3:

Rat liver lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 70 kDa,74 kDa

Observed band size: 110 kDa,50 kDa,72 kDa

false